Add all the components to a thin walled PCR tube (0.2ml thermo strip AB-0266, stores stocked, in 96well plate format) or 96 well plate and then spin in sigma 4K15 centrifuge @ 2000rpm for 10secs), cap tubes and LABEL THEM, spinning down should mix them.
Perform cycle sequencing reactions on a PCR machine MJ Tetrad close and tighten lid use heated lid) and run the following program: